Mouse Embryonic Stem Cells Expressing GDNF Show Enhanced Dopaminergic Differentiation and Promote Behavioral Recovery After Grafting in Parkinsonian Rats.
Mouse Embryonic Stem Cells Expressing GDNF Show Enhanced Dopaminergic Differentiation and Promote Behavioral Recovery After Grafting in Parkinsonian Rats.
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DOI：
10.3389/fcell.2021.661656
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发表时间：
2021
影响因子：
5.5
通讯作者：
Velasco I
中科院分区：
文献类型：
--
作者：
Lara-Rodarte R;Cortés D;Soriano K;Carmona F;Rocha L;Estudillo E;López-Ornelas A;Velasco I
帕金森病（Parkinson's disease，PD）是一种以中脑黑质多巴胺能神经元（DaNs）的进行性缺失和脑内多巴胺含量减少为特征的疾病。移植胚胎干细胞分化的DaN已被提出作为当前药物治疗的替代疗法。纹状体内移植从小鼠或人ESC分化的这种DaN改善运动性能，恢复DA释放，并抑制多巴胺受体超敏感性。然而，移植的神经元在大脑中存活的百分比很低。胶质细胞源性神经营养因子（GDNF）是DaNs的一种强存活因子。GDNF已被证明是神经营养的DaNs在体外和体内，并诱导轴突发芽和成熟。在这里，我们设计小鼠胚胎干细胞组成性产生人GDNF，分析DaN分化和可能的神经保护作用后，在体外毒性的挑战，或移植分化的DaN到帕金森病大鼠纹状体的转基因GDNF。GDNF在小鼠胚胎干细胞体外分化过程中的过表达增加了中脑DaNs的比例。这些转基因细胞在体外对6-羟基多巴胺的敏感性低于对照细胞。在移植对照或GDNF转基因DaNs后，我们观察到在药理学和非药理学行为测试中的显著恢复，以及纹状体DA释放增加，表明DaNs在脑中是功能性的。移植体积，存活神经元的数量，纹状体中存在的DaNs的数量，和移植物中DaNs的比例显着高于移植GDNF表达细胞的大鼠，与对照细胞相比。有趣的是，移植表达GDNF的细胞后，大鼠脑中没有发现形态学改变。这种方法是新颖的，因为先前的工作使用了DaN与表达GDNF的其他细胞类型的共移植，或者在DaN移植之前或之后在宿主组织中进行病毒转导。总之，GDNF生产小鼠胚胎干细胞有助于增强中脑分化，并允许在体外6-羟基多巴胺的挑战后，以及移植后在受损的纹状体更高数量的存活DaNs。这些表达GDNF的ESC可用于改善移植后的神经元存活。
Parkinson’s disease (PD) is characterized by the progressive loss of midbrain dopaminergic neurons (DaNs) of the substantia nigra pars compacta and the decrease of dopamine in the brain. Grafting DaN differentiated from embryonic stem cells (ESCs) has been proposed as an alternative therapy for current pharmacological treatments. Intrastriatal grafting of such DaNs differentiated from mouse or human ESCs improves motor performance, restores DA release, and suppresses dopamine receptor super-sensitivity. However, a low percentage of grafted neurons survive in the brain. Glial cell line-derived neurotrophic factor (GDNF) is a strong survival factor for DaNs. GDNF has proved to be neurotrophic for DaNs in vitro and in vivo, and induces axonal sprouting and maturation. Here, we engineered mouse ESCs to constitutively produce human GDNF, to analyze DaN differentiation and the possible neuroprotection by transgenic GDNF after toxic challenges in vitro, or after grafting differentiated DaNs into the striatum of Parkinsonian rats. GDNF overexpression throughout in vitro differentiation of mouse ESCs increases the proportion of midbrain DaNs. These transgenic cells were less sensitive than control cells to 6-hydroxydopamine in vitro. After grafting control or GDNF transgenic DaNs in hemi-Parkinsonian rats, we observed significant recoveries in both pharmacological and non-pharmacological behavioral tests, as well as increased striatal DA release, indicating that DaNs are functional in the brain. The graft volume, the number of surviving neurons, the number of DaNs present in the striatum, and the proportion of DaNs in the grafts were significantly higher in rats transplanted with GDNF-expressing cells, when compared to control cells. Interestingly, no morphological alterations in the brain of rats were found after grafting of GDNF-expressing cells. This approach is novel, because previous works have use co-grafting of DaNs with other cell types that express GDNF, or viral transduction in the host tissue before or after grafting of DaNs. In conclusion, GDNF production by mouse ESCs contributes to enhanced midbrain differentiation and permits a higher number of surviving DaNs after a 6-hydroxydopamine challenge in vitro, as well as post-grafting in the lesioned striatum. These GDNF-expressing ESCs can be useful to improve neuronal survival after transplantation.
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影响因子：
3.6
作者：
Das AT;Tenenbaum L;Berkhout B
通讯作者：
Berkhout B
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作者：
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通讯作者：
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